Liposomes enhance bioremediation of oil-contaminated soil

Liposomes (composed of soy phosphatides) in the form of small unilamellar vesicles (SUV), when added to soil contaminated by crude oil, accelerate bioremediation. After three weeks incubation at 30 degrees C, using soil experimentally contaminated (with 10,000 ppm crude oil), level of bioremediation increased from 40% without SUV to 75% with SUV (0.1 wt% phospholipids per dry weight soil). Similarly, for accidentally contaminated soil (with approximately 17,000 ppm crude oil), addition of 0.1 wt% SUV to the soil increased the bioremediation level from 55 to 80%. The enhancing effect of liposomes is explained by two interrelated phenomena: a large increase both in total bacteria number and in diversity of bacterial species in the soil. Comparison after four weeks revealed 21 bacterial species in the presence of liposomes (many being oil-degrading bacterial species) and only nine species in the absence of liposomes. Both effects may be related to the physical effects of liposome phospholipids, which modify the crude oil by wetting it, thereby making it more accessible to the microorganisms. In addition, liposome phospholipids serve as phosphate and nitrogen sources for the bacteria.     

Liposomes Enhance Bioremediation of Oil-Contaminated Soil

Abstract

Liposomes (composed of soy phosphatides) in the form of small unilamellar vesicles (SUV), when added to soil contaminated by crude oil, accelerate bioremediation. After three weeks incubation at 30°C, using soil experimentally contaminated (with 10,000 ppm crude oil), level of bioremediation increased from 40% without SUV to 75% with SUV (0.1 wt% phospholipids per dry weight soil). Similarly, for accidentally contaminated soil (with ～17,000 ppm crude oil), addition of 0.1 wt% SUV to the soil increased the bioremediation level from 55 to 80%. The enhancing effect of liposomes is explained by two interrelated phenomena: a large increase both in total bacteria number and in diversity of bacterial species in the soil. Comparison after four weeks revealed 21 bacterial species in the presence of liposomes (many being oil-degrading bacterial species) and only nine species in the absence of liposomes. Both effects may be related to the physical effects of liposome phospholipids, which modify the crude oil by wetting it, thereby making it more accessible to the microorganisms. In addition, liposome phospholipids serve as phosphate and nitrogen sources for the bacteria.     

An Intermediate-Scale Lysimeter Facility for Subsurface Bioremediation Research

A lysimeter facility at Oak Ridge National Laboratory, originally constructed to investigate leaching from low-level radioactive waste, was converted for use as an intermediate-scale facility for subsurface bioremediation research. The six experimental lysimeters are 2.5 m diameter by 4 m deep. The number and size of the lysimeters allow for replicate experiments and extensive sampling of the soil under controlled conditions. The facility provided containment of the contaminated soil, leachate, and microorganisms; positive control of the water table within the lysimeter; the ability to aerate the subsurface; multiple means of adding nutrients, electron acceptors, and electron donors to the subsurface; instrumentation for monitoring oxygen level, temperature, and moisture level; and means for obtaining samples of groundwater, soil, and liquid and gas samples from the soil pores. The flexibility of the facility allows for simulation of a wide range of subsurface bioremediation technologies. Startup and operational procedures and the advantages and disadvantages of the lysimeter facility are discussed. The facility is currently available to the bioremediation research community.     

高效石油降解菌群的构建及对芳香烃化合物萘的协同降解性能

【背景】石油作为一类混杂有机化合物,一旦产生污染就会对人类和环境造成严重的危害。【目的】从新疆石油污染土壤中分离筛选石油降解菌,为石油污染土壤的生物修复提供数据支持及技术参考。【方法】以石油为唯一碳源,通过富集培养、筛选分离得到123株单菌,根据菌落形态挑选出30个不同形态菌株,通过16S rRNA基因序列确定其种属,构建系统发育树;通过原油降解实验筛选出高效石油降解菌,以芳香烃的标志化合物萘为唯一碳源筛选出高效降解菌株,并分别筛选可降解水杨酸、邻苯二酚的菌株。【结果】分离筛选出5株高效石油降解菌,降解率高于85%;萘、水杨酸和邻苯二酚降解菌株各获得一株,将3种菌株按照1:1:1的接种比例对萘进行降解,萘的降解率从单菌60.74%提升到89.40%,菌株间的分工协作可以提高有机物的降解效率。【结论】筛选得到的菌株丰富了石油降解微生物菌种库,不同微生物菌株之间的分工协作为石油污染物的降解提供了新思路,为进一步研究石油污染治理提供参考。     

Degradation of crude oil in the rhizosphere of Sorghum bicolor

Dissipation of petroleum contaminants in the rhizosphere is likely the result of enhanced microbial degradation. Plant roots may encourage rhizosphere microbial activity through exudation of nutrients and by providing channels for increased water flow and gas diffusion. Phytoremediation of crude oil in soil was examined in this study using carefully selected plant species monitored over specific plant growth stages. Four sorghum (Sorghum bicolor L.) genotypes with differing root characteristics and levels of exudation were established in a sandy loam soil contaminated with 2700 mg crude oil/kg soil. Soils were sampled at three stages of plant growth: five leaf, flowering, and maturity. All vegetated treatments were associated with higher remediation efficiency, resulting in significantly lower total petroleum hydrocarbon concentrations than unvegetated controls. A relationship between root exudation and bioremediation efficiency was not apparent for these genotypes, although the presence of all sorghum genotypes resulted in significant removal of crude oil from the impacted soil.     

Enhanced crude oil biodegradation in soil via biostimulation

Research on feasible methods for the enhancement of bioremediation in soil contaminated by crude oil is vital in oil-exporting countries such as Kuwait, where crude oil is a major pollutant and the environment is hostile to biodegradation. This study investigated the possibility of enhancing crude oil bioremediation by supplementing soil with cost-effective organic materials derived from two widespread locally grown trees, Conocarpus and Tamarix. Amendments in soils increased the counts of soil microbiota by up to 98% and enhanced their activity by up to 95.5%. The increase in the biodegradation of crude oil (75%) and high levels of alkB expression substantiated the efficiency of the proposed amendment technology for the bioremediation of hydrocarbon-contaminated sites. The identification of crude-oil-degrading bacteria revealed the dominance of the genus Microbacterium (39.6%), Sphingopyxis soli (19.3%), and Bordetella petrii (19.6%) in unamended, Conocarpus-amended, and Tamarix-amended contaminated soils, respectively. Although soil amendments favored the growth of Gram-negative bacteria and reduced bacterial diversity, the structures of bacterial communities were not significantly altered.     

Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica

AIMS: To identify native Antarctic bacteria capable of oil degradation at low temperatures. METHODS AND RESULTS: Oil contaminated and pristine soils from Signy Island (South Orkney Islands, Antarctica) were examined for bacteria capable of oil degradation at low temperatures. Of the 300 isolates cultured, Pseudomonas strain ST41 grew on the widest range of hydrocarbons at 4 degrees C. ST41 was used in microcosm studies of low temperature bioremediation of oil-contaminated soils. Microcosm experiments showed that at 4 degrees C the levels of oil degradation increased, relative to the controls, with (i) the addition of ST41 to the existing soil microbial population (bioaugmentation), (ii) the addition of nutrients (biostimulation) and to the greatest extent with (iii) a combination of both treatments (bioaugmentation and biostimulation). Addition of water to oil contaminated soil (hydration) also enhanced oil degradation, although less than the other treatments. Analysis of the dominant species in the microcosms after 12 weeks, using temporal temperature gradient gel electrophoresis, showed Pseudomonas species to be the dominant soil bacteria in both bioaugmented and biostimulated microcosms. CONCLUSIONS: Addition of water and nutrients may enhance oil degradation through the biostimulation of indigenous oil-degrading microbial populations within the soil. However, bioaugmentation with Antarctic bacteria capable of efficient low temperature hydrocarbon degradation may enhance the rate of bioremediation if applied soon after the spill. SIGNIFICANCE AND IMPACT OF THE STUDY: In the future, native soil bacteria could be of use in bioremediation technologies in Antarctica.     

生物反应器法处理油泥污染土壤的研究

采油过程产生的油泥是整个石油烃污染源的重点。在陆地生态环境中 ,烃类的大量存在往往对植物的生物学质量产生不利影响 ,更重要的是石油中的一些多环芳烃是致癌和致突变物质 ,这些致癌和致突变的有机污染物进入农田生态系统后 ,在动植物体内逐渐富集 ,进而威胁人类的生存和健康[1 ,1 1 ] 。大量的废弃油泥 ,不仅污染农田 ,同时也给石油行业带来巨大的经济损失。污染土壤的治理主要有物理、化学和生物 (生物修复 )方法 ,生物修复方法被认为最有生命力。污染土壤生物修复技术主要有 3种 ,即原位处理、挖掘堆置处理和反应器处理。反应器处理是…     

石油烃污染土壤的生物修复

从中原油田污染土壤中通过实验室驯化培养分离到一组能以中原原油为碳源的快速生长的石油烃降解菌.用该组降解菌接种原油污染土壤,研究其原位生物联合修复实验,接种降解菌的各区分别种植大豆、施有机肥料、施有机肥料和锯末,与空白试样作对比.经过120d的联合修复,各区石油降解菌的总数(lgcfu/g)由接种时的5.25分别变为7.79、4.96、5.15、4.67.石油烃降解率分别达到89.4%、72.5%、76.7%、49.2%.表明分离的该组石油烃降解菌是一组高效降解菌且其与植物联合修复石油污染土壤能显著提高修复效果.     

Bioremediation of Crude Oil-Contaminated Soil Using Slurry-Phase Biological Treatment and Land Farming Techniques

Field-scale experiments on bioremediation of soil heavily contaminated with crude oil were undertaken on the territory of the Kokuyskoye oil field (Perm region, West Urals, Russia) owned by the LUKOIL Company. The pollution consisted of the contents of a oil waste storage pit, which mostly received soils contaminated after accidental oil spills and also the solid n-alkane (paraffin) wastes removed from the surface of drilling equipment. Laboratory analyses of soil samples indicated contamination levels up to 200?g/kg of total recoverable petroleum hydrocarbons (TRPH). Average oil composition consisted of 64% aliphatics, 25% aromatics, 8% heterocyclics, and 3% of tars/asphaltenes. Ex situ bioremediation techniques involved the successive treatment of contaminated soil using a bioslurry reactor and land farming cells. An oleophilic biofertilizer based on Rhodococcus surfactant complexes was used in both treatment systems. An aerobic slurry bioreactor was designed, and the biofertilizer applied weekly. Slurry-phase biotreatment of the contaminated soil resulted in an 88% reduction in oil concentration after 2 months. The resulting reactor product, containing approximately 25?g/kg of TRPH, was then loaded into land farming cells for further decontamination. To enhance bioremediation, different treatments (e.g., soil tilling, bulking with woodchips, watering, and biofertilizer addition) were used. The rates of oil biodegradation were 300 to 600?ppm/day. As a result, contamination levels dropped to 1.0 to 1.5?g/kg of TRPH after 5 to 7 weeks. Tertiary soil management involved phytoremediation where land farming cells were seeded with a mixture of three species of perennial grass. The effect of phytoremediation on the residual decontamination and rehabilitation of soil fertility is being evaluated.     

Effect of the tropical grass Brachiaria brizantha (Hochst. ex A. Rich.) Stapf on microbial population and activity in petroleum-contaminated soil

The effect of the tropical pasture grass Brachiaria brizantha on numbers of bacteria, fungi and degraders of alkanes, aromatics, cycloalkanes and crude oil in petroleum hydrocarbon contaminated and uncontaminated savannah soil was evaluated. Substrate induced soil respiration and soil pH were compared between planted and unplanted soil. B. brizantha had a mostly increasing effect on microbial numbers. As an exception, growth of bacteria was not or negatively affected. Microbial respiration and pH were always lower in planted than in unplanted soil. Low pH may result from enhanced oil degradation in planted soil leading to an accumulation of organic acids. A comparable stimulation of crude oil degraders and fungi in planted soil points to the importance of fungi. Since they tolerate lower pH values than bacteria, they are considered to play a central role in oil degradation. Given that the enhancement of crude oil degradation under the influence of B. brizantha could not clearly be correlated to microbial numbers and activity, other factors like oxygen availability, plant enzymes and synergistic degradation by microbial consortia have to be considered.     

Oil Palm Empty Fruit Bunch and Sugarcane Bagasse Enhance the Bioremediation of Soil Artificially Polluted by Crude Oil

Contamination of soil by petroleum hydrocarbons is becoming prevalent in Malaysia. Infiltration of soil contamination into groundwater poses a great threat to the ecosystem and human health. Bioremediation can occur naturally or can be enhanced with supplementation of microorganisms and fertilizers. However, fertilizers are expensive and therefore alternative nutrient-rich biomaterials are required. In this study, two organic wastes from agricultural industry (i.e., sugarcane bagasse and oil palm empty fruit bunch) were investigated for possible enhanced bioremediation of soil contaminated with Tapis crude oil. Two bacterial strains isolated and characterized previously (i.e., Pseudomonas aeruginosa UKMP-14T and Acinetobacter baumannii UKMP-12T) were used in this study. Sugarcane bagasse (5% and 15%, w/w) and oil palm empty fruit bunch (20%, w/w) were mixed with soil (500 g) spiked with Tapis crude oil (3%, v/w). The treated soils as well as controls were incubated for 20 days under controlled conditions. Sampling was carried out every four days to measure the number of bacterial colonies (CFU/g) and to determine the percentage of oil degradation by gas chromatography. The two biostimulating agents were able to maintain the soil moisture holding capacity, pH, and temperature at 38-40% volumetric moisture content (VMC), 7.0, and 29–30°C; respectively. The growth of bacteria consortium after 20 days in the treatment with sugarcane bagasse and oil palm empty fruit bunch had increased to 10.3 CFU/g and 9.5 CFU/g, respectively. The percentage of hydrocarbon degradation was higher in the soil amended with sugarcane bagasse (100%) when compared to that of oil palm empty fruit bunch (97%) after 20 days. Our results demonstrated the potential of sugarcane bagasse and oil palm empty fruit bunch as good substrates for enhanced bioremediation of soil contaminated with petroleum crude oil.     

Analysing the role of soil properties, initial biomass and ozone on observed plant growth variability in a lysimeter study

This simulation study is based on a lysimeter experiment with juvenile beech trees (Fagus sylvatica L.) which were grown under ambient or doubled ambient atmospheric ozone concentrations. The aim of the study was to analyze the role of differences in soil properties, differences in initial biomass and ozone impacts on observed plant growth variability at the eight lysimeters of this experiment. For this purpose, we established a new simulation model based on the model system Expert-N by coupling soil water and nitrogen transport models with the plant growth model PLATHO, which was already tested and applied for juvenile beech. In order to parameterize the soil model, for all lysimeters soil hydraulic parameters as well as carbon and nitrogen stocks were measured. Simulation results reveal that the observed decreased growth rates under elevated ozone are due to ozone impacts on plant growth, whereas the high plant growth variability between lysimeters is to a major part the consequence of differences in soil hydraulic properties. Differences in initial biomass are of minor importance to explain plant growth variability in this experiment.     

Oil desorption from mineral and organic materials using biosurfactant complexes produced by Rhodococcus species

Rhodococcus strains from the culture collection at the Institute of Ecology and Genetics of Microorganisms, Perm, Russia were examined for biosurfactant production during growth on n-alkanes and the ability to remove oil associated with contaminated sands and oil shale cuttings. Members of the genus, particularly R. ruber, were shown to produce low toxicity surfactants effective in removing oil from surfaces. The extent of desorption was inversely related to the concentration of high molecular weight hydrocarbons, namely asphaltenes and resins. In addition, crude surfactant complexes enhanced the degradation of crude oil, in the short term, when added to contaminated agricultural soil during bioremediation studies utilizing biopiling technology.     

Bioremediation of soil polluted with fuels by sequential multiple injection of native microorganisms: Field-scale processes in Poland

Research was conducted to estimate impact of the multiple bioaugmentation on the treatment of soil contaminated by fuels - diesel oil and aircraft fuel. The bacteria used to inoculate the remediation plots were isolated from the polluted soil and proliferated in field conditions. The amount of biomass applied to the polluted soil was set to ensure the total number of bacteria in soil 10⁷-10⁸ cfu/g d.w. The multiple inoculation of soil with indigenous bacteria active in diesel oil and engine oil (plot A) degradation increased bioremediation effectiveness by 50% in comparison to the non-inoculated control soil and by 30% in comparison to the soil that was inoculated only once. The multiple inoculation of soil with indigenous microorganisms was then applied in bioremediation of the soil polluted with double high concentration of diesel oil (soil B) and in bioremediation of the soil polluted with aircraft fuel (soil C). The process efficiency was 80% and 98% removal of TPH for soil B and C, respectively.     

The genome sequence of Polymorphum gilvum SL003B-26A1(T) reveals its genetic basis for crude oil degradation and adaptation to the saline soil

Polymorphum gilvum SL003B-26A1(T) is the type strain of a novel species in the recently published novel genus Polymorphum isolated from saline soil contaminated with crude oil. It is capable of using crude oil as the sole carbon and energy source and can adapt to saline soil at a temperature of 45°C. The Polymorphum gilvum genome provides a genetic basis for understanding how the strain could degrade crude oil and adapt to a saline environment. Genome analysis revealed the versatility of the strain for emulsifying crude oil, metabolizing aromatic compounds (a characteristic specific to the Polymorphum gilvum genome in comparison with other known genomes of oil-degrading bacteria), as well as possibly metabolizing n-alkanes through the LadA pathway. In addition, COG analysis revealed Polymorphum gilvum SL003B-26A1(T) has significantly higher abundances of the proteins responsible for cell motility, lipid transport and metabolism, and secondary metabolite biosynthesis, transport and catabolism than the average levels found in all other genomes sequenced thus far, but lower abundances of the proteins responsible for carbohydrate transport and metabolism, defense mechanisms, and translation than the average levels. These traits support the adaptability of Polymorphum gilvum to a crude oil-contaminated saline environment. The Polymorphum gilvum genome could serve as a platform for further study of oil-degrading microorganisms for bioremediation and microbial-enhanced oil recovery in harsh saline environments.     

石油污染土壤堆制微生物降解研究

采用异位生物修复技术堆式堆制处理方法 ,对辽河油田原油污染土壤进行了生物修复处理研究 .处理工程设 4个处理料堆单元 ,每个处理单元长 118.5cm ,宽 6 5 .5cm ,高 12 .5cm .研究结果表明 ,当进行处理的石油污染土壤中石油烃总量为 5 .2 2 g·10 0 g-1土时 ,利用黄孢原毛平革菌 (Phanerochaetechrysospori um) ,经过 5 5d的运行 ,石油烃总量去除率达 5 4.2 % .堆制处理中影响污染土壤石油烃总量生物降解的主要变化因子为污染土壤的O2 和CO2 含量、降解石油烃微生物的数量、污染土壤pH的变化 .通过监测这些数据的变化 ,可直接反映该工程的处理石油污染土壤的效果 .本处理工程采用定期通风措施 ,操作简单、运行费用低廉 ,为石油污染土壤生物修复实用化提供了一种简单易行的污染土壤清洁技术 .     

Oil bioremediation in a high and a low phosphorus soil

Phosphorus (P) content may influence bioremediation of soils contaminated with crude oil. A soil testing high in plant available P (Weswood, 194 mg P kg^?1 soil) and one testing low in plant available P (Lufkin, 2 mg P kg^?1 soil) were selected for laboratory experiments on oil biodegradation. Plant available P content was determined using acidified ammonium acetate at pH 4.2 as the soil extractant. Soils were amended with 3, 6, and 9% crude oil by weight and incubated for 120 d at 25°C. Treatments consisted of a factorial arrangement, with soil, N, P, and oil concentration as factors. Addition of P without N generally did not enhance biodegradation. Addition of N without P approximately tripled the quantity of oil degraded. Addition of P and N together did not increase biodegradation of oil more than addition of N alone when oil concentration was 3%. At 6 and 9% oil concentrations, CO₂ evolution increased for both soils by adding P and N together in comparison to adding N alone, and total petroleum hydrocarbon (TPH) bio‐degradation increased by 30% for the Weswood soil by 60 d and at least 25% for the Lufkin soil by 30 d. The quantity of plant‐available P or total P in soil was not very useful in predicting need for supplemental P. Addition of P to soil to enhance oil degradation was only beneficial for oil concentrations above 3% and the positive effect for higher concentrations was transitory.     

Impact of Oil-Spill Contamination on a Soil Bacterial Community: A 40-Year History of Rehabilitation in the Arava Valley

The present study evaluated the effect of crude oil contamination on a microbial community in hyper-arid soils. The Evrona Nature Reserve of Israel, situated in the Arava, was exposed twice in the last 40 years to petroleum-hydrocarbon-spill pollution. The first pollution event took place 40 years ago and was never treated, presenting a unique future time-point perspective to the second (2014) contamination event. Soil samples were collected after the second spill, and abiotic properties and bacterial diversity in the sampled soil were analyzed. The results showed that there is a significant decrease over time in the number of observed bacterial species in the contaminated samples, coupled together with bacterial species replacement toward species capable of using source oil as the sole carbon and energy source. The presence of petroleum in soil significantly changed the composition and functional diversity of a microbial community, and the Evrona Nature Reserve is still in the middle of a bioremediation process even 40 years after the crude oil contamination.     

植物-微生物联合修复毒死蜱污染的土壤

本试验以毒死蜱污染土壤为研究材料,利用降解菌DSP-A分别与高丹草、紫花苜蓿、多花黑麦草进行联合修复,探讨了植物-微生物联合修复毒死蜱污染土壤的效果,以及影响联合修复的因素,结果表明,植物．微生物联合修复的效果优于单一的植物修复及单一的微生物修复效果。与DSP—A菌群较合适的植物是高丹草,该组合对毒死蜱的降解率达到96．44％,其次是多花黑麦草。研究了微生物数量、植株密度以及土壤湿度对联合修复效果的影响,结果表明,DSP．A菌菌液稀释倍数越大,联合修复的效果越差。植株密度对联合修复的影响,主要表现为对植物根系生长的影响。植株密度越大,对生存环境的竞争越激烈,植物根系的生长越不好。除了紫花苜蓿外,高丹草和多花黑麦草根系的生长均受到影响。高丹草种植密度为12株／盆时,与DSP—A菌的联合修复效果最好,多花黑麦草则为10株／盆。土壤湿度是影响联合修复的重要因素,不仅影响植物的生长,对微生物的生长也有影响。土壤湿度过大,造成土壤的含氧量降低,不利于植物根系和好氧细菌的生长,从而影响土壤中农药的降解。土壤湿度过小,容易造成植株缺水,根系生长和微生物的生长。高丹草与DSP．A菌、多花黑麦草与DSP—A菌联合修复最适浇水量都为20mL／d,紫花苜蓿与DSP—A菌联合修复最适浇水量都为15mL／d。